NDLI: Expression and Antibody Production of Japanese Encephalitis Virus RNA Polymerase (NS5) Protein

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Expression and Antibody Production of Japanese Encephalitis Virus RNA Polymerase (NS5) Protein

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Expression and Antibody Production of Japanese Encephalitis Virus RNA Polymerase (NS5) Protein

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Kim, J. M. Yun, S. I. Song, B. H. Choi, Y. J. Park, J. S. Lee, Y. M.
Description	Responsible library: WPRO
Abstract	Japanese encephalitis virus (JEV), a member of mosquito-borne flaviviruses, is the leading cause of viral encephalitis in a large geographic area of Southeast Asia and Australia. JEV contains a single-stranded positive-sense RNA genome, which encodes its own RNA-dependent RNA polymerase (NS5) that is required for genomic RNA replication. In this study, we have described a pair of mouse antisera specific to the N- or C-terminal region of the NS5. Initially, two hydrophilic regions corresponding to the N-terminus and C-terminus of the NS5 protein were individually amplified by reverse transcription-PCR from the genomic RNA of JEV K87P39 strain. The amplified DNA fragments were cloned into a prokaryotic expression vector, pGEX-4T-1; the resulting constructs were used for the expression of GST fusion proteins, designated GST/NS5N and GST/NS5C, in E. coli BL-21 strain. Following immunization of three BALB/c mice with each of the purified GST/NS5N and GST/NS5C, we obtained two pools of the antisera, specifically recognizing the ~103-kDa NS5 and several smaller NS5-related proteins in BHK-21 and Vero cells infected with JEV K87P39 strain. Overall, we have successfully expressed the N- and C-terminal regions of JEV NS5 fused to the C-terminus of GST and generated the mouse antisera capable of recognizing the NS5 and its related proteins in JEV-infected cells. This would provide a valuable reagent for the study of JEV NS5 in the viral life cycle.
File Format	HTM / HTML
ISSN	15982467
e-ISSN	20930429
Journal	Journal of Bacteriology and Virology
Issue Number	1
Volume Number	39
Language	Korean
Publisher	Korean Society for Microbiology Korean Society of Virology
Publisher Date	2009-01-01
Publisher Place	Korea (South)
Access Restriction	Open
Subject Keyword	Discipline Microbiology Discipline Virology Discipline Infectious Diseases Discipline Bacteriology Gst Gene Carbon Ion Proteins Upper Case En
Content Type	Text
Resource Type	Article
Subject	Virology Immunology Microbiology

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